TWI245771B - Micro-region selectively activating hydrophobic chip and its preparing method - Google Patents

Abstract

This invention is related to provide a micro-region selectively activating hydrophobic chip/microarray. The preparing method is related to use a hydrophobic material and a compound having one functional group protected by a protective group to prepare a hydrophobic polymer by means of mixing, branching or polymerizing. Among which, the functional group is related to imide or cyclic amide, the protective group is related to photo acid group like tosyloxy group. And then apply the hydrophobic polymer on a base. After treating the hydrophobic polymer with selectively optical activation, the polymer micro-region having functional activity is formed and is separated from the non-activated polymer. The chip of this invention is suitable for preparing biochip/microarray with high density and high efficiency.

Description

1245771 V. Description of the invention (1) Description of the invention Field of the invention: The present invention relates to a sa / microarray for selective activation of microdomains and a method for preparing the same. The present invention is particularly related to-a sacrilic polymer prepared by mixing, grafting or polymerizing a raw material and a raw material with a functional group protected by a protective group. The hydrophobic polymer is coated on a substrate, and then selectively activated by selective light to form a hydrophobic wafer that is selectively activated by a micro-region. Phenol tablets are suitable for the preparation of hydrophobic microarrays with selective activation of microdomains. Known technology ... In the current biochip and biomicroarray technology, most of the preparation methods involve silanizing the substrate surface (3 丨 1 & 11 丨 2 to 011), and then interacting with the biomaterial.联 反应 (crossiinking reaction). During the silylation process, the substrate surface is activated according to the material of the substrate surface, and then treated with a hydrophilic silane, such as amino-propyl-tetraethoxy-siiane (APTES). The next cross-linking reaction is to fix the biological material on the substrate by a cross-linking agent (such as glutaraldehyde). The disadvantages of this method include: it is only applicable to certain specific substrates, the reaction time is long, the reaction efficiency is low, and the low activity of the fixed biological material. Moreover, the covalently bonded surface prepared by this method is hydrophilic, and cross-contamination between micro-regions is easily generated when preparing a selective microarray. U.S. Patent No. 5,837,8 60 and World Patent No. WO 98/39481

0648-5793TWF (N); 89-19; ch i umeow.ptd Page 4 1245771 V. Description of the invention (2) · The nucleic acid probe is fixed on a hydrophobic silane, such as mercapto-s i iane ) Technology on glass or silicon wafers. In this method, a hydrophobic thiol group ((Hs__), me reap to-group) is covalently fixed on a substrate. The hydrophobicity of the thiol group is suitable for high-density, immobilized nucleic acids / oligonucleic acids. . However, this method must modify the biological material so that it has a thiol group, but this modification reaction is not easy to perform and has low efficiency. The cost is relatively high. The thiol-modified nucleic acid probe is placed in a thiol. A disulfide bond is formed between the silane-modified biological material and the substrate surface to complete the immobilization of the nucleic acid probe. Blanchar, Αρ · etal · (Bisensens and Bi〇electronics, 1996, 11 (6/7): 687-69〇) disclosed coating a photoresist on a substrate, and then using a microelectrochemical (microelectrochemical) -electromechanical) mask is developed to form the hydrophilic points of the micro-regions, and the areas outside the hydrophilic points are hydrophobic. After treatment, the surface of the tritium can be subjected to high-density nucleic acid probe micro-arrays and synchronous (insi) processes. The conventional technology does not disclose the preparation of a hydrophobic polymer and—a functional group with a protective group (such as a fluorenimine or a cyclic amine group) —combined, grafted, or polymerized into a hydrophobic polymer. Thing.

The application of the hydrophobic polymer prepared by coating it on an organic or organic substrate to form a micro-area light-activated wafer is summarized. Therefore, the main f η „Lp of the present invention, such as One of the main goals of King Al is to provide a hydrophobic chip for selective activation of micro-locusts, wall pendulums, and sterilization 4 WU £ domains suitable for high-density microarrays. this

1245771 V. Description of the invention (3) The wafer of the invention includes a substrate, and the substrate is coated with a hydrophobic polymer layer. The hydrophobic polymer layer includes functionally activated polymer microdomains separated by inactive polymers. The functionally activated polymer includes a hydrophobic substance and a compound having a functional group, wherein the functional group is an imide or a cyclic amide. The non-reactive polymer includes a hydrophobic substance and a compound with a functional group protected by a protective group, wherein the functional group is a fluorimide or a cyclic amine compound, and the protective group is a photo acid group. . A photoacid group is a chemical molecule that is sensitive to light of a specific wavelength. Under the irradiation of light of this specific wavelength, the chemical between the photoacid group and the original covalently bonded atom will gradually break and detach. A covalently bonded atom of an acid group and a hydrogen atom create a new bond. Photoacid groups are called hydrogen atoms because they generate hydrogen atoms after reacting with light. The reaction is similar to ordinary acidic substances. Examples of suitable photoacid groups are sulfonyloxy group, N-methanesulfonyloxy group, N-trifluoro-methanesulfonyloxy group gr〇up), camphor sul fonyl oxy group, or tos 1 ο xygroup. Another object of the present invention is to provide a microdomain selective activation. The hydrophobic microarray includes a substrate, and the substrate is coated with a hydrophobic polymer layer. The hydrophobic polymer layer includes functionally activated polymer microdomains separated by inactive polymers. Does the above functional activated polysaccharide contain a hydrophobic property? And a compound with a functional group, of which: can make: imine or cyclic amino compounds. Inactive polymer contains a hydrophobic substance

0648-5793TWF (N); 89-19; chiumeow.ptd 1245771

$ And a compound with a functional group protected by a protective group, in which the B-b group is a fluorenimine or a cyclic amine compound, and the protective group is another basic purpose of the basic invention of a photoacid to provide a microdomain selection A method for preparing a saponified activated water-soluble wafer. The preparation method includes the following steps: First, a hydrophobic polymer is dissolved in a solvent to form a hydrophobic polymer solution; and then the hydrophobic polymer solution is coated. On a substrate (such as a 2 substrate or an inorganic substrate); removing the above-mentioned solvents; selective photo-active activation of the hydrophobic polymer, forming a functionally active polymer between inactive polymers Micro-domains, polymer micro-domains are separated by active polymers. Detailed description of the invention One of the characteristics of the present invention is a preparation method, which comprises the following solvent to form a solvate solution and apply it to a base to remove the above solvent; active for polymerization and functional polymerization in non-active polymers The polymer segment is prepared according to the present invention with a protective group and protected polymerization to form a microregion selective activated hydrophobic wafer. First, a hydrophobic polymer is dissolved in an aqueous polymer solution. The hydrophobic polymer substrate (such as an organic substrate or an inorganic substrate); the water-based polymer layer is subjected to selective photoreceptive actives (that is, the above-mentioned hydrophobic polymer) to form micro-regions among the polymer micro-regions. The hydrophobic polymer is a compound composed of a hydrophobic material and a functional group, and the compound can be a combination of a cyclic imine or a cyclic amine by mixing, grafting, or sulfonyl.

1245771 V. Description of the invention (5) The protective group is a photoacid group, such as sul f ony loxy group, N-methanesulfonyloxy group, N- N-trifluoro-methanesulfonyloxy group, N-trifluoro-methanesulfonyloxy group, camphorsulfonyloxy group or tosloxy gr0Up, among which 703 ~ 1. } ^ Groups are more suitable. Then, the hydrophobic polymer is coated on a substrate made of organic or inorganic material, and the solvent is removed to form a hydrophobic layer. The hydrophobic polymer has a covalent bond functional group protected by a protective group. Due to the large volume of protective groups, this hydrophobic sound is relatively inert. Stomach & In the photosensitivity activation step, the irradiation of a certain area (or dot) line is used to activate the functional group of >. When applied to an array of biomaterials, the biojunction can be achieved without the distance between the field of the activated polymer of the present invention and the biological material of each other. The present invention has a step of a hydrophobic layer. Hydrophobic polymerization polymers are inert and difficult to mask with a mask. Materials can only be distinguished from micro-domains. The benefits are greatly reduced and mixed, so the functional activation can therefore improve the exposed functions and the polymer can react with the functional inactive energy. Use a photomask while avoiding a substantial increase in polymer production. High production photomask on the hydrophobic layer. Protective groups on specific substrates migrate to form biomaterials that retain their protective groups and therefore 'use this crystal to activate polymer micro-regional polymer reactions. Polymeric microdomains and photoactive activation technology avoid the need to fix different microarray densities and bond efficiencies of biomaterials. Moreover, the light of the work wavelength is divided. The obtained valence bond is maintained and the sheet is prepared to form microdomains. There is a gap between each of the microdomains and the microdomains. The junction reaction can only be activated.

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The functional groups contained in the polymer microdomain are densely functional groups (such as fluorenimine or cyclic amine group): two changes to contain protected materials in mixed, grafted or polymerized derivatives and hydrophobic another Features are adjusted for .2 ratio. Material or modified affinity: When the reaction = (for euPnile), the combination produces another:: =… the hydrophilicity of the strong micro area. Areas that have not been activated = are hydrophobic surfaces, and the activated areas react with biological materials to form hydrophilic microdomains, which helps to distinguish between activated and unactivated areas. This feature is conducive to the subsequent biochemical reaction.

The term "enhanced microregion hydrophilicity" means that the amidine or cyclic amine structure in the hydrophobic layer prepared by the above method is attacked by a nucleophilic group (such as an amine-modified oligonucleotide probe), Through ring-opening reaction, another hydrophilic group is generated, which leads to the strengthening of the hydrophilicity of the micro-region. At this time, the originally hydrophobic surface is converted to hydrophilic, while the other regions remain hydrophobic. When using a protective hydrophobic polymer is poly (styrene-co- [N- (p-toluenesulfonyloxy) maleenediimide]) (po1y (styrene-co- [N- ( toxy1oxy) ma1eimide])), when the functional hydrophobic polymer is PS M I, the reaction is shown in Reaction Formula I:

Reaction Formula I:

0648-5793TWF (N); 89-19; chiumeow.ptd page 9 1245771 V. Description of the invention (7)

P〇ly (s iyiene-c ΰ- [N-(tos yloxy) male imide]) Poly (s tyr ene-c <?-KialeiKiide) biologi cal m aterials

Unless otherwise stated, technical and scientific terms used herein are the same as those familiar with the field of the invention. The abbreviations used in this manual have the following meanings: APTES, amino-propyl-tetraethoxy-siliane; PS, polystyrene; MI, maleimide maleimide); PSMI, poly (styrene-co-maleimide); PE, (polyethylene); PEM I, poly (ethylene-co-maleimide) Poly (ethylene-co-maleimide); Ts

0648-5793TWF (N); 89-19; ch i umeow.ptd page 10 1245771 i. Description of invention (8) tosyloxy group; TsCl, toluenesulfonic acid chloride; TsOMI, N- ( P-toluene xanthanyloxy) maleimide (N- (tosyloxy) maleimide); IPA, isopropyl alcohol; ΑΙΒΝ, N, Ν'-azobisisobutyronitrile ( The chemical formula of the N, N, -azobisisobutyronitrile) ° Ts group is shown below:

The selectively activated hydrophobic 'can be organic or organic. Organic materials can be based on monomers such as acrylate. The monomers include, but do not need a substrate. If necessary, a polymer based on a base. For example, the technical composition of the present invention having a hydrophobic surface under pressure molding includes, but is not limited to, a polymer that is not limited to the substrate material and can be used for coating a hydrophobic substrate selected according to the present invention. . Suitable are esters, acrylics, acrylic acrylates. Suitable baseless porcelain, glass or metal. Such as PEMI polymers. In addition, a photoacid group is used as a protection, and a chemical wafer is prepared according to a preferred implementation or pressure molding. A suitable method for preparing a hydrophobic polymer using an inorganic base wafer, an inorganic material, that is, the method is to synthesize olefin, styrene, acryl, acryl-based acrylic acid, or acryl-based acrylic from silicon-based monomers. Pottery can include PSMI or bottom. It can also contain water-based polymers. Micro-regions of the projecting surface can be highly hydrophilic. Those skilled in the art are familiar with the limitation.

〇648-5793T1VF (N); 89-I9; ch1umeow. Page 11 1245771 V. Description of the invention (9) The bottom surface is activated to strengthen the bonding force between the hydrophobic polymer and the substrate surface. The activation step includes treating the surface of the substrate with an acid or an alkali, or performing plasma activation treatment. According to the preparation method of the present invention, before coating the hydrophobic polymer, the surface of the substrate may be subjected to a cleaning procedure to prevent impurities from contaminating or contaminating the substrate surface. This cleaning step is based on the characteristics of the substrate material, and is cleaned with a solvent and / or son i cat ion. Suitable solvents include, but are not limited to, surfactants, water, alcohols, or acetone. The hydrophobic polymer of the present invention is prepared by mixing, grafting, or polymerizing a hydrophobic material and a compound with a functional group protected by a protective group. The functional group is imide (imide ) Or cyclic amide; protective group is photo acid group, such as sulfonyloxy group, N-methanesulfonyloxy group, N- N-trif luoro-methanesulfonyloxy group, camphor sul fonyl oxy group, and tosloxy group, of which tosyloxy Groups are more suitable. The hydrophobic polymer of the present invention may include any compound mixed, grafted, or polymerized with fluorenimine or cyclic amine groups, such as 4-amino-phthalimide ) Or 3-methyl-2-pyridone. Such hydrophobic materials include acetophenone, urethane, ethylene, or derivatives thereof.发明 的 _ In the preferred embodiment, the hydrophobic material used is polystyrene or ethylene The resulting hydrophobic polymer comprises P S ΜI or Ρ Ε Μ I.

1245771 The technology of coating a hydrophobic polymer on a substrate is unlimited, and is well known to those skilled in the chemical and semiconductor process related fields, including spin coating, screen printing (scree η printing, roller coating, rolling coating, curtain coating, or dip coating, and the like. In a preferred embodiment, the coating of the hydrophobic polymer is achieved by spin coating, preferably spin coating at 3,000 to 6,000 rpm, and more preferably spin coating at 4,000 rpm. After the coating of the hydrophobic polymer on the substrate is completed, the excess solvent is removed, for example, by vacuum evaporation, heating evaporation, or evaporation under reduced pressure. Among them, if heating and evaporation are performed, the temperature should be used to avoid damaging the substrate or unnecessary polymerization. After the solvent is removed, the wafer of the present invention can be subsequently prepared. The hydrophobic polymer then coated on the substrate will undergo selective photoactivation. In the present invention, the protecting group is a photoacid, such as a sulfonyloxy group, a N-methanesulfonyloxy group, and a N-trifluoro-methanesulfonyloxy group ( N_trifluoro-methanesulfonyloxy group), camphorsulfonyloxy group or tosloxy group, among which the tosyloxy group is more suitable, and the protected function in the hydrophobic polymer The reactivity of groups to biomaterials is very low. In the present invention, the wavelength of the light suitable for the photoreactive activation reaction is 248 nm or 312 nm. Appropriate photoprogramming procedures allow the conforminyl group in the hydrophobic polymer to be removed 'from the functional group, thereby activating the functional group in the hydrophobic polymer. After this step, the activated functional group can form with the biological material

1245771 V. Description of the invention (11) Covalent bonding According to a preferred embodiment of the present invention, the area or point to be activated is defined by a photomask during the activation of the light display. Multiple sets of photomasks can be applied to different biomaterials on the same wafer to form a microarray, which means 'first use a photomask to activate the area preset to fix the first biomaterial'. A biomaterial is fixed in these activated areas 'the area activated by the second photomask 7 is activated with a second photomask' to fix the second biomaterial in the activated area of the second photomask 'So on and so forth. One of the advantages of this selective photoactive activation wafer is that after being activated, the functionally active regions are separated by hydrophobic polymers that are not activated (that is, protected by the protection group 4), thus allowing the biological material cloth to be greatly improved. The density placed on the microarray and the application of the photomask also help the production of high density microarrays. M Another feature of the present invention is to provide a microdomain selective activated hydrophobic 1 * 4 array. The test array contains a bioactive material, which is fixed on a hydrophobic wafer selectively activated by a micro-region as described above. T 1 The fixation of the above bioactive material is through biomolecules and I domain selection of the present invention. Achieved by contact activation of hydrophobic wafers. The bioactive material 7 s has a nucleophilic group, such as a monoamine group, which exists in the fraction or chemically modified form of the bioactive material. The bond between the nucleophilic group in the biologically active material and the functional group in the functional / tonated polymer is formed through a ring-opening reaction. ^ f The bioactive materials used in the present invention include the following biomolecules: nucleic acids, nucleic acids, peptide nucleic acids (PNA), antigens, antibodies, enzymes, or eggs

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White matter. This type of biomolecule is related to the present invention, μ t # 官 & its ^ = T domain selectively activates hydrophobic linkage), and then the bioactive material is fixed on the wafer, 1 (.) In the conventional technology, the bioactive material and the wafer The connection is achieved through two + step = reactions. First, a silane-based polymerization reaction is performed, and then a parent linker such as glutaric acid is added.) The method of the present invention will μ ^ ^ ^ ^ ftVb /-should effectiveness. The time is reduced by 7 π and 77. Another feature of the present invention is the dynamic conversion of hydrophobicity / hydrophilicity of the microdomain when the biomaterial is fixed on the wafer. High-density microarrays can be made based on water-repellent functional layers. In subsequent reactions, the positioning of biological materials can be enhanced by hydrophilizing the microregions. In addition, according to the preparation method of the present invention, the preparation time of the microarray will be greatly shortened compared with the traditional manufacturing method, and at the same time, the wafer prepared according to the preparation method of the present invention has better homogeneity. This means that there will be an average functional group on the substrate (ie fluorenimine or cyclic amine group) and bond strength. In addition, the hydrophobic microarray for selective activation of the microdomains of the present invention greatly shortens the time required to fix the biological material on the wafer. In a preferred embodiment, the time required for fixation is less than 40 minutes. The spirit of the present invention will be further described in the following examples, but the examples are not intended to limit the present invention. Example 1: Preparation of TsOM I. 0.5 g of N-hydroxymaleimide and 0.5 g of TsCl were mixed in 50 mi of toluene.

0648-5793TWF (N); 89-19; chiumeow.ptd

1245771 V. Description of the invention (13) (toluene) / IPA solution, heated to 60. 〇, and allowed to react under a nitrogen atmosphere for 3 hours' to obtain a T s 0 Μ I solution. Example 2: Preparation of poly (styrene-co— [ν- (p-toluenesulfonyloxy) maleimidoimide]) Preparation of poly (styrene—C0— [N— (p-toluenesulfonyl The reaction of poly (styrene-co- [N- (tosyloxy) maleimide])) is shown in the reaction formula II:

I I HC = CH2

OTs

Fbly (stynenB-i »-[N < tosyb3y > tiaJeiitiide])

TsOMI

Slyreng added polyethylene and TS 0 MI in a ratio of 4.1 mole ratio to a nitrogen or argon reaction bottle containing toluene, added 0.5% AIBN dropwise, and carried out a copolymerization reaction at 60 C for two hours. Then, the reaction was terminated by aeration to obtain a toluene solution of poly (styrene-co- [N- (p-a-benzylsulfonyloxy) maleimidoimide]). Example 3: Poly (styrene-co_ [N- (p-toluenesulfonyloxy) cis-butene di

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The immobilized effect of acylimine]) will be based on the poly (styrene-co- [N- (p-toluenesulfonyl) maleimide)] prepared in Example 2) (poly (styrene-co- [N- (tosyloxy) maleimide])) toluene solution 'and spin-coated on a glass wafer at 4,000 rpm to form a poly (styrene-co- [N- (pair Tosyloxy) maleicimide]) wafer. The wafer was dried in an oven at 1000 ° C to remove toluene. Take a synthetic oligo probe, which has fluorescein on the 5 end and amine modification on the 3 'end. This nucleic acid probe is placed on the polymer modified wafer to fix the oligo nucleic acid probe.化 反应。 Reaction. The conditions for the immobilization reaction were: · 0.5 probe was dissolved in 2X SSC buffer (pH 7.0) and placed on the wafer ', and allowed to react at 37 ° C for 1 hour; then 0.2% SDS Clean the wafer for 10 minutes. Cleaned and unwashed (control group) wafers were analyzed for their effect on solidification by fluorescence analysis. The results are shown in Figure 1 (unwashed 10, cleaned 1 2), showing very few nucleic acid probes successfully fixed On a poly (styrene-co- [N- (p-toluenesulfonyloxy) maleimidoimide]) wafer. Example 4: PSMI oligodeoxynucleotide immobilization effect According to Example 3, the present ethylene-co- [n- (p-toluene xanthoyloxy) maleimide imide]] & (poly (styrene- co-[N-Uosyloxyhaieimide])) wafer. After the toluene was removed from the wafer, the wafer was exposed to light at a wavelength of 2 48 nm for J minutes, and the poly (styrene_co- [N- (p-toluenesulfonyloxy) maleimide) Amine]) into PSMI. Then, according to the method described in Example 3,

1245771 V. Description of the invention (15) The m-needle is placed on the wafer, and the oligo nucleic acid (control group) wafer is determined by Yingguang analysis; ί: Please refer to Figure 2 (unwashed 20, washed) The number of oligonucleotide probes fixed on the PSMI chip is significantly higher than that of Example 3. Example 5: Hybridization reaction of specific oligos The chip used in this example is the same as that described in Example 4. This example uses The nucleic acid is MO (containing 29 bases, 5 with 15 thymidine and amine groups), 〇3 (containing 29 bases, 5 without amine groups at the ends), (containing a nucleic acid, 5 'end There is an amine group), and the needle used for the flag hybridization reaction is a complementary sequence of these nucleic acid-raising nucleic acids, and the 3 end has a fluorescent flag. The solidification conditions were 2X SSC, pH 7.0; 3X SSC, pH 7.0.0; 10x SSC, pH 3.0. The immobilization reaction time was i hours. The hybridization reaction lasted 4 hours. Figure 3 shows the results of judging the hybridization reaction by fluorescence analysis, showing wafers 30 cleaned with AlO3 probes at 2X SSC, PH7; wafers 32 cleaned with ai03 probes at 3x SSC, PH7; Wafer 34 cleaned with A10 03 probe at 10χ SSC, ρΗ3; Wafer 36 cleaned with 03 probe at 2X §sc, ρΗ7; Wafer 38 cleaned with 03 probe at 3x ssc, ρΗ7 ; Wafer 40 cleaned with 03 probe, i0x ssc, ρΗ3; Wafer 42 cleaned with ps probe '2x SSC, ρΗ7; Wafer 44 cleaned with P3 probe, 3x SSC, ρΗ7; used ρ3 probe, wafer 46 cleaned at 10x ssc, ρ3.

0648-5793TWF (N); 89-19; chiumeow.ptd Page 18 1245771 V. Description of the invention (16) The functionally activated polymers of the present invention, that is, PSMI and PEMI retain functional groups (such as imine d) It is bonded to biological materials, so biological materials such as amine-modified DNA can be stably fixed on the wafer surface. In Figure 1, due to the functional part of poly (benzylethylene-co- [N- (p-tolueneoxy) maleimidoimide]), a huge photoacid group (p-toluenesulfonyl) (Oxygen) protected, most of the nucleic acid probes failed to form a bond with the polymer on the wafer and were washed away in subsequent cleaning steps. In contrast, poly (styrene-C0- [N_ (p-toluenesulfonyloxy) enediylimide]) has been converted to PSMI (see Figure 2), The acid needle still maintained a fixation rate of 90% after cleaning with 0.2% SDS. Figure 3 shows the efficacy of three different sets of probes on light-activated wafers. AIO3 needle contains 29 test bases, 15 thymidine and amine groups at the 5 'end,% probe contains "bases have amine groups, P3 probe contains 29 bases, 5, right end ms With a fixed base under pH 3.0 and pH 7.0 environment, there is a wide base. Each of these probes is observed, and the 03 probe is activated on the wafer because of the lack of 5, and the result shows 'this-light display'. The fixed rate of the activated wafer table X. The nucleic acid probe has better reactivity and can modify the accuracy of the oligodity. The adsorption of the surface of the wafer improves the function of the m-chip in the present invention. Degree, can be prepared by changing the functional group contained in the hydrophobic domain dense functional groups (such as fluorenimine or cyclic amine group): Heart = 0648-5793TWF (N) on page 19; 89 -19; ch i umeow. ptd 1245771 V. Description of the invention 〇7) —___ Proportion of the material to be adjusted. 觭 荖 所 人 夕 — In addition, the fixed efficiency is also related to the density increase of the Hg 2 group energy group is not good. Because Γ Too much hydrophilicity is secret to the preparation of high-density microarrays. Therefore, the Guan Saqi 00 paint sound library in the hydrophobic polymer layer depends on different conditions. The adjustment is another important factor for the preparation of the 4 degree hydrogel group. * The sparsity of the present invention. 成份 The composition of the water-soluble polymer that has a good homogeneity after being coated on the substrate is not limited to molybdenum. ， 成 / 取 # For example, u polyethylene test: 1 ethylene / polystyrene. For example, hydrophilic polymer ^ &a; the main chain of the polymer can also be used in micro-regions to strengthen the wafer. Any hydrophobic polymer that is official ^. /, And contains at least one imine or sphingosine group can be applied to the present invention. The invention has been disclosed above by way of example, but it is not intended to be limiting. 5 ^ But you can be familiar with this skill without departing from the spirit of the present invention: application and retouching, so the scope of protection of the present invention shall be determined by the scope of the application # patent scope of this case shall prevail. See also 0648-5793TWF ( N); 89-19; chiumeow.ptd Page 20 1245771 Schematic illustrations Simple illustrations: The spirit and advantages of the present invention will be further explained in more detail by the following and the illustrations, where the illustrations are: 1 The picture shows a coating of poly (styrene-co- [N- (p-toluenesulfonyloxy) cis Butenedioyl imine]) (0〇17 (3 七 丫 ^ 1 ^-(: 〇-[^-(1: 〇371〇 \ 7) 11 ^ 1 至 11 ^ (16))) and not Fluorescent intensity display of an oligo nucleic acid probe with a fluorescent marker on a glass wafer activated by light display. The second figure is a poly (styrene-co- [N- (p-toluenesulfonate) coated with Acyloxy) maleimide]) (P〇ly (styrene-co- [N- (tosyloxy) maleimide])) and a light-activated glass wafer with a fluorescent marker Fluorescence intensity display after nucleic acid probe. Figure 3 shows the fluorescence intensity display of three groups of different oligonucleotide probes hybridized with their respective nucleic acid targets. The nucleic acid-producing probes are fixed on a poly (styrene-co- [N- ( P-toluenesulfonyloxy) maleenediimide]) (Poly (styrene-co- [N- (tosy lOxy) maleimide])) and activated by photo-optical activation on a glass wafer. Explanation of symbols: 1 0 ~ unwashed, 1 2 ~ washed; 2 0 ~ unwashed; 2 2 ~ washed;

0648-5793TWF (N); 89-19; chiumeow.ptd P.21 1245771 The diagram briefly explains the product of 30 ^ aio3 Probe 9 washed with 2 x SSC :, pH7; 32, ^ Al〇3 Probe 9 with 3 x SSC :, pH 7 was cleaned; 34 ^ AlO3 probe was washed with 1 Ox SSC, pH3; 36 '"〇3 probe was washed with 2x SSC, p H 7; 38" "3 probe 9 was washed with 3x SSC, p H 7 cleaning, 40, 〇3 probe 9 was washed with 10x SSC, pH3; 42 ', P3 probe 5 was washed with 2x SSC, pH7; 44 ,, P3 probe? washed with 3x SSC, pH7, 46, P3 probe 9 was washed with 10x SSC, pH3.

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Claims (1)

1245771 Case No. 91122291 \, please patent patent park I. A microregion selective activation of a hydrophobic wafer, including if IL supplementation ......... a hydrophobic polymer layer, a polymer segment of a micro-region and a material with a base and a water system. The hydrophobic material and the functional group are protected. Group, N-sulfenylsulfonylsulfonyloxy group is coated on the substrate, the hydrophobic layer contains functional activation, the functional activated polymer micro-regions are unactivated, wherein the functional activated polymer comprises a A compound of a hydrophobic material group, the unactivated polymer contains a compound with a functional group protected by a protective group, and a photoacid group; the material contains styrene, amino potassium ester, ethylene or a derivative thereof The group is an acid imine or a cyclic amino group, and the photoacid group includes a continylfluorenyloxy group, an N-trifluoro-fluorenylsulfonyloxy group, a camphor group, or a p-toluenesulfonyloxy group. 2. The micro-domain selectively activated hydrophobic wafer as described in item 1 of the scope of the patent application, wherein the functionally activated polymer comprises poly (styrene-co-maleimidoimide), or poly ( Ethylene-co-maleimide). 3. The hydrophobic wafer with selective activation of the upper region as described in item 1 of the scope of the patent application, wherein the unactivated polymer comprises poly (styrene-co- [N- (p-ammoniumsulfonyloxy)) Maleimidoimide]), or poly (ethylene-co- [N- (p-benzylsulfonyloxy) maleimidoimide]). 4. The micro-region selectively activated hydrophobic wafer according to item 1 of the scope of the patent application, wherein the substrate comprises a polymer obtained by polymerizing an organic monomer, I P.23 1245771 Case No. 91 122291 Amendment C, claim patent scope Acrylic Where the organic monomer includes ethylene, styrene, propylene, ester, acrylate, alkyl acrylic acid or alkyl acrylate. 5. The microregion-selectively activated hydrophobic wafer as described in item 1 of the scope of the patent application, wherein the substrate comprises a poly (ethylene-C0-maleimidoimide) or poly (ethylene-C0 -Polymers of maleimide). 6. The micro-region selectively activated hydrophobic wafer according to item 1 of the scope of the patent application, wherein the substrate comprises an inorganic material, and the inorganic material comprises silicon crystal, ceramic, glass or metal. 7. The micro-region selectively activated hydrophobic wafer according to item 1 of the patent application scope, wherein the substrate further comprises a polymer having a photoacid group. 8. A hydrophobic microarray with selective activation of microdomains, comprising a substrate; a hydrophobic layer coated on the substrate, the hydrophobic layer comprising functionally activated polymer microdomains, It is separated by an unactivated polymer, wherein the functional activated polymer includes a hydrophobic material and a compound having a functional group, and the unactivated polymer includes a hydrophobic material and a protective group. A functional group of compounds, and the protecting group is a photoacid group, wherein the hydrophobic material comprises styrene, amino potassium acid, ethyl or its derivative, and the functional group is fluorene or imine Cyclic amine group, the photoacid group contains sulfonium Page 24 ζ 月 口 stated #, more) Original revision JJ _, ___ 1245771 Case No. 91122291 _ί: VI. Patent application group, N-methanesulfonyloxy group, N-trifluoromethanesulfonyl group An oxygen group, a camphor sulfonyloxy group or a p-toluenesulfonyloxy group; and a bioactive material containing an amine group, which includes a nucleophilic group, and the bioactive material passes through the nucleophilic amine group A ring-opening reaction between a group and a functional group of the functional activated polymer is fixed on the functional activated polymer. 9. The selectively activated hydrophobic microarray according to item 8 of the scope of the patent application, wherein the functionally activated polymer 7 comprises poly (styrene-C0-maleimidoimide), Or poly (ethylene-C0-maleicimide). -10. The hydrophobic microarray with selectively activated microdomains as described in item 8 of the scope of patent application, wherein the unactivated polymer comprises poly (styrene-co- [N- (p-toluenesulfonyloxy) ) Maleimidoimide]), or poly (ethylene-co- [N- (p-toluenesulfonyloxy) maleimidoimide]). 1 1. The hydrophobic microarray with selective activation of microdomains as described in item 8 of the scope of patent application, wherein the substrate comprises a polymer obtained by polymerizing an organic monomer, wherein the organic monomer comprises ethylene and styrene , Propylene, ester, acrylic acid, acrylic acid S purpose, acrylic acid or acrylic acid purpose. 1 2. The hydrophobic microarray with selective activation of microdomains as described in item 8 of the scope of the patent application, wherein the substrate comprises a substrate containing a poly (styrene-co-maleic acid) Amine) or poly (ethylene-co-cis-butyl) Page 25 2005. 06. 08. 009 1245771: ^ p 〇1 __ case number 91122291_ year. · Yuexin 1st __ VI, please patent scope of the polymer 13. The water-repellent microarray with selectively activated microdomains as described in item 8 of the scope of the patent application, wherein the substrate comprises an inorganic material, and the inorganic material comprises a silicon wafer, ceramic, glass, or metal. 14. The hydrophobic microarray with selectively activated microdomains as described in item 8 of the scope of the patent application, wherein the substrate further comprises a polymer having a photoacid group. 15. The hydrophobic microarray for selective activation of a microdomain as described in item 8 of the scope of the patent application, wherein the bioactive material comprises a nucleic acid, an oligonucleotide, a peptide nucleic acid, an antigen, an antibody, an enzyme, or a protein. 16. A method for preparing a hydrophobic wafer with microdomain selective activation, comprising the following steps: A hydrophobic polymer is dissolved in a solvent to obtain a hydrophobic polymer solution. The hydrophobic polymer is composed of a hydrophobic material and a compound having a functional group protected by a protective group, and is mixed and grafted. Prepared by polymerization or polymerization, the protecting group is a photoacid group, wherein the hydrophobic material comprises stupid ethylene, amino potassium potassium ester, ethylene or a derivative thereof, and the functional group is fluorenimine or a cyclic amino group The photoacid group includes a continuous acid group, an N-formic acidoxy group, an N-trifluoro-fluorenylsulfonyloxy group, a camphor sulfonyloxy group, or a p-sulfonylsulfonyloxy group. group; Page 26 1245771 Case No. 91122291 9 丨 3 years old, month said _ amendment :, please ask the patent fan garden to apply the hydrophobic polymer solution on a substrate; eliminate the solvent; and the hydrophobicity is carried out by light The selectively photo-active polymer of the hydrophobic polymer layer is converted into a functional activated polymer. 17. The method for preparing a hydrophobic wafer with selective activation of microdomains as described in item 16 of the scope of the patent application, further comprising using a photomask on the hydrophobic polymer during the photosensitivity activation, wherein The photomask has a pattern that covers areas that are not to be activated by the light display and exposes areas that are not to be activated by the light display. region. 18. The method for preparing a microdomain-selectively activated hydrophobic wafer as described in item 16 of the scope of patent application, wherein the hydrophobic polymer comprises poly (styrene-c0- [N- (p-toluenesulfonyl) (Oxy) maleimidoimide]), or poly (ethylene-co- [N- (p-phenylenesulfonyloxy) maleimidoimide]). 19. The method for preparing a microdomain selective activated hydrophobic wafer as described in item 16 of the scope of patent application, wherein the functionalized polymer comprises a polymer (Stupid Ethyl-C 0 -cisbutadiene diimide), or Poly (Ethylene-C 0 -cis-butyral diimide). 20. The method for preparing a microdomain selectively activated hydrophobic wafer as described in item 16 of the scope of the patent application, wherein the substrate comprises a polymer obtained by polymerizing an organic monomer, wherein the organic monomer comprises ethylene and benzene Ethylene, Propylene Page 27, 1245771, Case No. 91122291, Amendment VI, claim patent scope olefin, ester, acrylic acid, acrylate, alkyl acrylic acid or alkyl acrylic acid 2 1. Preparation of micro-region selection as described in item 16 of the scope of patent application. Method for hydrophobically activated hydrophobic wafers, wherein the substrate comprises a poly (styrene-C 0 -maleimidoimide) or poly (ethylene-C 0 -maleimidoimide) Of polymers. 22. The method for preparing a microregion-selectively activated hydrophobic wafer according to item 16 of the patent application scope, wherein the substrate comprises an inorganic material, and the inorganic material comprises a silicon wafer, ceramic, glass, or metal. 23. The method for preparing a microdomain-selectively activated hydrophobic wafer as described in item 16 of the patent application scope, further comprising activating the substrate before the step of coating the hydrophobic polymer solution. 24. The method for preparing a hydrophobic wafer with selective microdomain activation as described in item 23 of the scope of patent application, wherein the step of activating the substrate comprises treating the surface of the substrate with an acid or an alkali, or plasma activation. 2 5. The method for preparing a microdomain selective activated hydrophobic wafer as described in item 16 of the scope of patent application, further comprising cleaning the substrate before the step of applying the 'hydrophobic polymer solution'. Page 28, 1245771, Case No. 91122291 May, Inclined Amendment 6. Patent scope 26. The method for preparing hydrophobic wafers with selective activation of microdomains as described in item 25 of the patent application scope, wherein the cleaning step is Clean with a solvent and / or super-hybrid shock, the solvent containing an interfacial activator, water, alcohols or acetone 2 7. The method for preparing a hydrophobic microchip selective activation as described in item 16 of the scope of patent application The coating step includes spin coating, screen printing, roller coating, curtain coating, or dip coating technology. 2 8. The method for preparing a microregion-selectively activated hydrophobic wafer as described in item 16 of the scope of patent application, wherein the removal of the solvent is performed by vacuum evaporation, heating evaporation, or evaporation under reduced pressure. Page 29